Interrogating the function of motile ciliated cells in spinal curvature using a zebrafish model of adolescent idiopathic scoliosis

摘要 青少年特发性脊柱侧凸(AIS)是一种普遍的发育性疾病(影响约3%的 人群)来源不明，以出现异常的三维脊柱弯曲为特征 在青春期的各个阶段。最近，许多人工免疫系统的斑马鱼模型已经成为强大的 用于解决这种情况的病因学和表征基本生物学机制的工具 促进脊椎伸直。然而，这些模型的不同遗传基础，它们的多样性 脊柱的曲度和生长脊柱的复杂性--一种为脊柱的组织提供界面的组织 骨骼、肌肉和神经系统--意味着进行定量、表型分析的能力 关于微型计算机断层扫描(MicroCT)的数据是我们描述 调节和保持脊柱伸直。这项提案的目标是解决这一障碍，以 将一种新的定量、多尺度表型组学分析方法应用于斑马鱼AIS模型的研究进展， 这将使我们能够解决有关调节和 保持脊柱直立。 我的导师最近的研究表明，斑马鱼的突变会使运动纤毛在 幼年生长窗发育出三维脊柱曲度，概括了AIS。我们的中环 假说认为椎管室管膜的活动纤毛是局部的关键信号装置。 “感知”和脊柱直度和曲度的机械矫正。我们打算挑战这些 研究目的：(1)测试脊柱伸直时活动纤毛的空间要求。 (2)确定哪些特定的运动性纤毛细胞类型对直脊椎至关重要。(3)测试需求 活动纤毛下游机械轴上接触脑脊液的神经元。研究设计：我们将使用我们的 定量、多尺度表型组学平台，用于评估涉及以下两种操作之一的基因操作的影响 脊柱伸直时纤毛运动的整体、部分或细胞型特异性失活。这将通知我们的 了解活动的纤毛如何将信息传递到它们所在的组织环境，有助于我们的 基本了解脊柱如何“知道”如何变直，并为我们未来的方向提供信息 AIS的研究和治疗。

Summary Adolescent Idiopathic Scoliosis (AIS) is a prevalent developmental condition (affecting ~3% of the population) of unknown origin, characterized by abnormal, three-dimensional spinal curvatures that occur during phases of adolescent growth. Recently, numerous zebrafish models of AIS have emerged as powerful tools for addressing etiology of the condition and characterizing the fundamental biological mechanisms that facilitate spinal straightness. However, the varied genetic underpinnings of these models, the diversity of their spinal curvatures, and complexity of the growing spine – a tissue that provides an interface for tissues of the skeletal, muscular, and nervous systems – means that an ability to perform quantitative, phenotypic analysis on micro-computed tomography (microCT) data is a critical barrier to our ability to describe the pathways that mediate and maintain spinal straightness. The objectives of this proposal are to address this barrier to progress by applying a new modality of quantitative, multiscale phenomics analysis to zebrafish models of AIS, which will allow us to address fundamental questions about the developmental mechanisms that mediate and maintain spinal straightness. Recent work by my advisor has shown that zebrafish with mutations that paralyze motile cilia during windows of juvenile growth develop three-dimensional spinal curvatures recapitulating AIS. Our Central Hypothesis is that motile cilia of the spinal canal ependyma are critical signaling devices in the local `perception' and mechanical correction of spinal straightness and curvature. We intend to challenge these ideas through three Specific Aims: (1) Test the spatial requirement for motile cilia during spinal straightness. (2) Identify which specific motile ciliated cell types are critical for a straight spine. (3) Test the requirement for CSF-contacting neurons in a mechanical axis downstream of motile cilia. Research Design: We will use our quantitative, multiscale phenomics platform to evaluate the impact of genetic manipulations involving either global, partial, or cell-type specific inactivation of cilia motility on spinal straightness. This will inform our understanding of how motile cilia relay information to their local tissue environments, contribute to our fundamental understanding of how the spine “knows” to grow straight, and inform our future directions in the research and treatment of AIS.